Effectene Transfection Reagent is a nonliposomal lipid reagent for DNA transfection into a broad range of cell types. Due to its low cytotoxicity, Effectene Transfection Reagent is highly suitable for transfection of primary cells and many sensitive cell lines.

The Effectene Transfection Reagent is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

High transfection efficiencies using Effectene Reagent.

Transfection efficiencies using Effectene Reagent and two commonly used lipid-based reagents were compared. Murine teratocarcinoma F9 cells (5 x 105) were transfected in 6-well plates with a luciferase-reporter plasmid using optimized conditions based on the manufacturer's instructions for each reagent. Transfection efficiencies were determined by measuring luciferase activity 48 hours post-transfection, and are given as relative light units (RLU). (Data kindly provided by I. Clavereau, D. Petitprez, and I. Van Seuningen, Unité INSERM 377, Place de Verdun, Lille Cedex, France.)

Serum and DNA quantity vs. transfection efficiency.

The influence of serum and DNA quantity on transfection using Effectene Reagent was examined. COS-7 cells (2 x 104 per well) were seeded in 96-well plates one day before transfection. Cells were transfected using 0.01–1.0 µg beta-galactosidase-reporter plasmid and 0.08–8.0 µl Enhancer (DNA: Enhancer ratio of 1:8) and 2 µl Effectene Reagent, in either the presence or absence of serum. Each bar represents the average efficiency from 4 replicates 48 hours post-transfection.

Effectene transfection procedure.

DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 2-5 minutes. Effectene Reagent is then added and the mixture is incubated for 5-10 minutes to allow Effectene-DNA complexes to form. The complexes are mixed with growth medium (which can contain serum and antibiotics), and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression.

The application of recombinant DNA technology to fields such as drug discovery and development has led to an increased need for high-throughput transfection. Transfection using Effectene Transfection Reagent requires low amounts of DNA and minimal handling. In addition, removal of transfection complexes is not required, making this reagent highly suitable for high-throughput screening. Effectene Transfection Reagent is available in bulk quantities.

Principle

Effectene Transfection Reagent is an innovative non-liposomal lipid formulation that is used in conjunction with a special DNA-condensing enhancer and optimized buffer to achieve high transfection efficiencies. The enhancer first condenses the DNA molecules and Effectene Reagent subsequently coats them with cationic lipids providing a particularly efficient way of transferring DNA into eukaryotic cells. This feature ensures excellent reproducibility of transfection complex formation.

Procedure

The Effectene procedure has two steps. DNA is first mixed with Enhancer and a buffer that provides optimal salt conditions for efficient DNA condensation. This step requires just 2–5 minutes. Effectene Reagent is then added and the mixture is incubated for 5–10 minutes to allow Effectene–DNA complexes to form. The complexes are mixed with growth medium (which can contain serum and antibiotics), and added directly to the cells. The cells are then incubated until harvested and analyzed for gene expression (see flowchart "Effectene transfection procedure").

Applications

Effectene Transfection Reagent is suitable for transient and stable transfection of a broad range of cell types.

Features

Specifications

Applications

Plasmid transfection, protein overexpression, reporter studies

Cell type

Eukaryotic cells (primary cells and sensitive cells)

Controls

Not included

Features

Non-liposomal lipid formulation, minimal cytotoxicity

Nucleic acid

DNA

Number of possible transfections

160 transfections in 12-well plates / 1 ml reagent

Technology

Non-liposomal lipid formulation in conjonction with a DNA-condensing enhancer

The following protocol is optimized for transient transfection of 293 cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The following protocol is optimized for transient transfection of CHO cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.

The following protocol is optimized for transient transfection of COS-7 cells in 96-well plates without pre-plating of cells 24 h prior to transfection. Cell plating and transfection are performed on the same day, making this protocol rapid and convenient.